Cant, SH;
Pitcher, JA;
(2005)
G protein-coupled receptor kinase 2-mediated phosphorylation of ezrin is required for G protein-coupled receptor-dependent reorganization of the actin cytoskeleton.
MOL BIOL CELL
, 16
(7)
3088 - 3099.
10.1091/mbc.E04-10-0877.
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Abstract
G protein-coupled receptor kinase 2 (GRK2) phosphorylates and desensitizes activated G protein-coupled receptors (GPCRs). Here, we identify ezrin as a novel non-GPCR substrate of GRK2. GRK2 phosphorylates glutathione S-transferase (GST)-ezrin, but not an ezrin fusion protein lacking threonine 567 (T567), in vitro. These results suggest that T567, the regulatory phosphorylation site responsible for maintaining ezrin in its active conformation, represents the principle site of GRK2-mediated phosphorylation. Two lines of evidence indicate that GRK2-mediated ezrin-radixin-moesin (ERM) phosphorylation serves to link GPCR activation to cytoskeletal reorganization. First, in Hep2 cells muscarinic M1 receptor (M1MR) activation causes membrane ruffling. This ruffling response is ERM dependent and is accompanied by ERM phosphorylation. Inhibition of GRK2, but not rho kinase or protein kinase C, prevents ERM phosphorylation and membrane ruffling. Second, agonist-induced internalization of the beta(2)-adrenergic receptor (beta(2)AR) and M1MR is accompanied by ERM phosphorylation and localization of phosphorylated ERM to receptor-containing endocytic vesicles. The colocalization of internalized beta(2)AR and phosphorylated ERM is not dependent on Na+/H+ exchanger regulatory factor binding to the beta(2)AR. Inhibition of ezrin function impedes beta(2)AR internalization, further linking GPCR activation, GRK activity, and ezrin function. Overall, our results suggest that GRK2 serves not only to attenuate but also to transduce GPCR-mediated signals.
| Type: | Article |
|---|---|
| Title: | G protein-coupled receptor kinase 2-mediated phosphorylation of ezrin is required for G protein-coupled receptor-dependent reorganization of the actin cytoskeleton |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1091/mbc.E04-10-0877 |
| Publisher version: | http://www.ncbi.nlm.nih.gov/pmc/ articles/PMC11653... |
| Keywords: | EXCHANGER REGULATORY FACTOR, EZRIN/RADIXIN/MOESIN ERM PROTEINS, EPIDERMAL-GROWTH-FACTOR, AMINO-TERMINAL DOMAIN, BETA(2)-ADRENERGIC RECEPTOR, EPITHELIAL-CELLS, HOMOLOGY DOMAIN, BINDING-SITE, IN-VITRO, ACTIVATION |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL |
| URI: | https://discovery.ucl.ac.uk/id/eprint/9718 |
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